EP1536753A1

EP1536753A1 - Method for protecting and for modulating dermal-epidermal junctions

Info

Publication number: EP1536753A1
Application number: EP03798892A
Authority: EP
Inventors: Christine Jeanmaire; Gilles Pauly
Original assignee: Cognis France SAS
Current assignee: BASF Beauty Care Solutions France SAS
Priority date: 2002-09-13
Filing date: 2003-09-04
Publication date: 2005-06-08
Anticipated expiration: 2023-09-04
Also published as: US20060051302A1; KR20050049493A; EP1536753B1; EP1398019A1; JP2006507257A; WO2004030639A1; ES2569056T3

Abstract

The invention relates to a method for cosmetically treating and improving and/or protecting dermal-epidermal junctions of the skin, scalp and mucous membranes. This method is characterized in that a preparation containing at least one substance, which effects a modulation of plectin/HD1 and/or entactin/nidogen and/or perlecan is topically applied. The invention also relates to the use of a substance, which effects a modulation of plectin/HD1 and/or entactin/nidogen and/or perlecan, for producing cosmetic agents for improving dermal-epidermal junctions of the skin, scalp and mucous membranes, to the use of this substance for producing cosmetic agents for protecting against aging of the skin and oxidative stress, and to the use of this substance for producing cosmetic agents for protecting against damaging influences of environmental toxicants and UV radiation.

Description

Method for the protection and modulation of dermal epidermal junctions

Field of the invention

The invention is in the field of cosmetic preparations and relates to a cosmetic method for improving and / or protecting the dermal-epidermal junctions of the skin, scalp and mucosa and for protecting the human skin against aging, oxidative stress and harmful influences Environmental toxins and UN radiation. Furthermore, the invention relates to the use of a substance which effects a modulation of plectin / HDI and / or entactin / Νidogen and / or perlecan for the preparation of cosmetic agents for the improvement and / or protection of the dermal epidermal junctions.

State of the art

The basement membrane is a connecting cell structure between morphologically different tissues. In the skin it is essentially tissue surrounding blood vessels and between Dennis and epidermis. The latter region separates the epidermis and its appendages from the Dennis and is accordingly called the dermal epidermal junction (DEJ). The DEJ has a complex structure consisting of hemidesmosomes, intermediate filaments, anchoring filaments, lamina densa, and anchoring fibrils. Among the major biochemical components therein are laminin-5 in lamina lucida; the antigens AgBP 230 and AgPB180 as well as plectin / HDI in hemidesmosomes; Entactin / Νidogen and the proteoglycan Perlecan in the lamina lucida and lamina densa; Collagen type IN in the lamina densa and the proteoglycan collagen type NU as part of the anchoring fibrils in the sub-lamina densa. These components form an interacting network.

The DEJ represents the most important structure of the skin. It provides the connection between epidermis and underlying Dennis and preserves the integrity of the epithelial tissue by anchoring cells with the extracellular matrix via the special connecting cell complexes of hemi-desmosomes, filaments and fibrils.

On the one hand, it is a filter for the flow of specific molecules, on the other hand, it allows the exchange of information - for example about growth factors - between keratinocytes and Dennis. Age-related changes in the DEJ include the decreasing thickness of the junctions and the reduction of basal keratinocyte cytoplasmic cell villi in Dennis. The resulting decrease in surface area of the DEJ results in reduced tissue resistance, a decrease in skin tightening, and increased wrinkling. Other signs of aging such as doubling of the lamina densa, aging of anchoring fibrils, or modifications of DEJ cell components also cause anchoring via the epidermal system to become more relaxed with increasing age. The decrease in type VJJ collagen caused by aging is also explained by the hydrolysis of the collagen via metalloproteinases. Type NU collagen appears to be less resistant to proteases with age.

Damage induced by UV radiation, which leads to a reduction in the content of type NU collagen in the anchoring fibrils, results in a relaxation of the bond between Dennis and epidermis and a consequent decrease in skin elasticity and increase in wrinkles.

Many diseases that affect the DEJ lead to the development of subepi- malmal growths. They show as a common feature a reduced cohesion between Dennis and epidermis, which manifests itself in the formation of indentations at the DEJ level.

With the aim of improving the function of the dermal epidermal junctions, DEJ components in particular, such as lamin (FR 2813018, WO 97/48415), or kalinin (WO 92/17498), have been used in cosmetic and dermatological formulations. Also, the stimulation of type IV collagen by magnesium aspartate (WO 99/62481), saponins (FR 2779058) or plant extracts (EP 668072) and the stimulation of type Nu collagen by Elaginsäure (WO 99/16415), plant extract of Potentilla erecta (WO 98 / 19664) or Bertholletia extract (US 6004568) have been disclosed for the preparation of cosmetic preparations against aging, wrinkling and tightening of the skin.

Nevertheless, there is still a need for effective protection of the skin against environmental aging. The object of the present patent application has therefore been to find new mechanisms for improving the dermal-epidermal junctions of the skin, scalp and mucous membrane, which delay skin aging and protect the skin, scalp and mucous membrane against environmental influences, oxidative stress, contribute toxic substances or UV radiation and thus can be used effectively in cosmetic preparations for topical application. Description of the invention

The invention relates to methods for the cosmetic treatment for the improvement and / or protection of the dermal-epidermal junctions of the skin, scalp and mucous membrane, characterized in that a preparation containing at least one substance which has a modulation of Plectin / HDI and / or Entactin / Nidogen and / or Perlecan effected, topically applied.

Further objects of the invention are the use of a substance which effects a modulation of plectin / HDI and / or entactin / nidogen and / or pearlcan for the preparation of cosmetic agents for the improvement and / or protection of the dermal-epidermal junctions of the skin, scalp and Mucosa, the use of this substance for the production of cosmetic agents for protection against aging of the skin and the use of the substance for the production of cosmetic agents for protection against oxidative stress, harmful effects of environmental toxins and UN radiation.

Surprisingly, it has been found that the modulation of molecules such as plectin / HDI, entactin / Νidogen and / or pearlcan leads to the preservation and improvement of the dermal-epidermal junctions of the skin, scalp and mucous membrane. The function of the DEJ is an essential requirement not only for health but also for cosmetics. It ensures a good cohesion between epidermis and the underlying tissues of Dennis, thus preserving the elasticity and firmness of the skin and prevents the formation of wrinkles. In addition, the DEJ ensures good skin care through the passage of vital molecules between the epidermis and Dennis, while also providing protection against the penetration of damaging molecules into deeper layers of the skin.

The modulation of plectin / HDI, entactin / Νidogen and / or pearlcan via the topical application of a preparation which stimulates these substances has now shown that in this way the dermal epidermal junction resp. the entire complex network of the DEJ can be strengthened. This results in a tightening of the skin and reduction of wrinkles. The improved anchoring between the components of the DEJ and the associated increased stability and increasing elasticity of the tissue can effectively prevent aging phenomena, even when they are caused by UV radiation.

Due to the improvement of the molecular function, the exchange between keratinocytes and Dennis and the nutrition of the skin is optimized, thus protecting not only the skin against aging, but also against damaging influences of UN radiation and toxic environmental influences the defense against damaging molecules is strengthened. The DEJ surrounds the hair follicles on the scalp and also provides protection for the follicles, so that strengthening the DEJ in this area leads to an improvement in the hair's properties and, in particular, is effective against hair loss or hair damage.

Thus, the topically applied cosmetic compositions which contain at least one substance which effects the modulation of plectin / HDI and / or entactin / nidogen and / or perlecan have a preventive effect against skin aging and damaging influences by oxidative stress, environmental toxins and UV radiation on skin, scalp, hair and mucous membrane. However, the modulation of the special molecules leads not only to the preventive effect but also to an accelerated regeneration of the skin, scalp and mucous membrane after an already occurred damage.

As modulators, plant extracts, in particular the extract of Pisum sativum, Ruscus aculeatus, Centella asiatica, Calendula officinalis, Aesculus Hippocastanum, and / or Hibiscus esculentus have proven to be suitable. However, it is also possible to modulate plectin / HDI and / or entactin / nidogen and / or pearlcan via the administration of low molecular weight peptides which are required for the synthesis of the DEJ components and have a sequence similar to components of plectin / HDI and / or Entactin / nidogen and / or pearlcan.

Also observed was a modulation of the molecules via at least one substance selected from the group consisting of mannitol, cyclodextrin, yeast extract, panthenol, propylene glycol, ammonium glycyrrhizate and disodium succinate. In particular, the combination of these ingredients contributes to a beneficial effect on the dermal epidermal junctions.

Since the DEJ is highly susceptible to degradation by a variety of different proteases, the use of plant extracts, peptides, and other agents with anti-protease activity contribute significantly to preserving the structure. Agents with anti-protease activity in combination with substances that cause the modulation of plectin / HDI and / or entactin / nidogen and / or pearlcan may contribute to an above-average increase in efficacy.

In addition to these substances or plant extracts, the cosmetics may also contain UV sun protection factors and / or antioxidants. The combination of substances which induce a modulation of plectin / HDI and / or entactin / nidogen and / or pearlcane with UV sun protection factors and / or antioxidants leads, through the different mechanisms, to a synergistic mode of action and offers outstanding protection against damaging influences and skin aging by UV light. Plectin / HDI

Plectin and HD1 are synonyms for the same molecule. It is localized in the hemi-desmosomes, has a molecular mass of about 500 daltons and serves to anchor proteins of the cytoskeleton such as keratin, vimentin or proteins of the microtubules.

Entactin / Nido gene

Entactin and nidogen are also different terms for the same molecule. It represents a glycoprotein with a molecular mass of about 150 daltons, which consists of two terminal globular regions linked by a long-chain structure. This glycoprotein is responsible for the structural stability of the DEJ by providing a link between laminin and type IV collagen and anchoring a variety of other components, such as fibulin or perlecan.

perlecan

All basement membranes contain proteoglycans. The proteoglycan, which occurs mainly in DEJ, is Perlecan, a heparan sulfate synthesized by dermal fibroblasts. Perlecan consists of a large protein core and three heparan sulfate chains. Its task is, among other things, to stabilize a bond between laminin-6 and nidogen. In addition to anchoring functions, heparan sulfate proteoglycans also bind diffusing molecules such as enzymes and growth factors and may also affect cell behavior and properties.

UV sunscreen and antioxidants

Under UV sun protection factors, for example, at room temperature, liquid or crystalline organic substances (sunscreen) to understand that are able to absorb ultraviolet rays and the absorbed energy in the form of longer-wave radiation, e.g. Heat again. UVB filters can be oil-soluble or water-soluble. As oil-soluble substances are e.g. to call:

3-Benzylidencampher or 3-Benzylidennorcampher and its derivatives, for example 3- (4-methylbenzylidene) camphor; 4-aminobenzoic acid derivatives, preferably 2-ethylhexyl 4- (dimethylamino) benzoate, 2-octyl 4- (dimethylamino) benzoate and 4 ~ (dimethylamino) benzoic acid amyl ester; Esters of cinnamic acid, preferably 4-methoxycinnamic acid 2-ethylhexyl ester, propyl 4-methoxycinnamate, isoamyl 4-methoxycinnamate 2-cyano-3,3-phenylcinnamic acid 2-ethylhexyl ester (octocrylene);

Esters of salicylic acid, preferably 2-ethylhexyl salicylate, 4-isopropylbenzyl salicylate, homomenthyl salicylate; Derivatives of benzophenone, preferably 2-hydroxy-4-methoxybenzophenone, 2-hydroxy-4-methoxy-4'-methylbenzophenone, 2,2'-dihydroxy-4-methoxybenzophenone;

Esters of benzalmalonic acid, preferably di-2-ethylhexyl 4-methoxybenzmalonate; Triazine derivatives, e.g. 2,4,6-trianilino (p-carbo-2'-ethyl-hexyloxy) -l, 3,5-triazine and octyl triazone, or dioctyl butamido triazone (Uvasorb® HEB);

> Propane-1,3-diones, such as e.g. l- (4-tert-butylphenyl) -3- (4'methoxyphenyl) propane-l, 3-dione;

> Ketotricyclo (5.2.1.0) decane derivatives.

Suitable water-soluble substances are:

2-phenylbenzimidazole-5-sulfonic acid and its alkali, alkaline earth, ammonium, alkylammonium, alkanolammonium and glucammonium salts; Sulfonic acid derivatives of benzophenones, preferably 2-hydroxy-4-methoxybenzophenone-5-sulfonic acid and its salts;

Sulfonic acid derivatives of the 3-benzylidene camphor, e.g. 4- (2-oxo-3-bornylidenemethyl) benzenesulfonic acid and 2-methyl-5- (2-oxo-3-bomylidene) sulfonic acid and salts thereof.

As a typical UV-A filter in particular derivatives of benzoylmethane are suitable, such as, for example, 1- (4'-tert-butylphenyl) -3- (4'-methoxyphenyl) propan-1, 3-dione, 4-tert-butyl 4'-methoxydibenzoylmethane (Parsol® 1789), 1-phenyl-3- (4'-isopropylphenyl) -propane-1, 3-dione and enamine compounds. Of course, the UV-A and UV-B filters can also be used in mixtures. Particularly favorable combinations consist of the derivatives of benzoylmethane, e.g. 4-tert-butyl-4'-methoxydibenzoylmethane (Parsol® 1789) and 2-cyano-3,3-phenylcinnamic acid 2-ethylhexyl ester (octocrylene) in combination with esters of cinnamic acid, preferably 2-ethylhexyl 4-methoxycinnamate and / or 4-methoxycinnamic acid propyl ester and / or 4-methoxycinnamic acid isoamyl ester. Advantageously, such combinations are made with water-soluble filters, e.g. 2-phenylbenzimidazole-5-sulfonic acid and their alkali, alkaline earth, ammonium, Alkyla monium-, Alkanolammonium- and Glucammoniumsalze combined.

In addition to the soluble substances mentioned, insoluble photoprotective pigments, namely finely dispersed metal oxides or salts, are also suitable for this purpose. Examples of suitable In particular, metal oxides are zinc oxide and titanium dioxide and, in addition, oxides of iron, zirconium, silicon, manganese, aluminum and cerium, and mixtures thereof. As salts silicates (talc), barium sulfate or zinc stearate can be used. The oxides and salts are used in the form of the pigments for skin-care and skin-protecting emulsions and decorative cosmetics. The particles should have an average diameter of less than 100 nm, preferably between 5 and 50 μm and in particular between 15 and 30 nm. They may have a spherical shape, but it is also possible to use those particles which have an ellipsoidal or otherwise deviating shape from the spherical shape. The pigments may also be surface-treated, ie hydrophilized or hydrophobicized. Typical examples are coated titanium dioxides, such as titanium dioxide T 805 (Degussa) or Eusolex® T2000 (Merck). Suitable hydrophobic coating agents are in particular silicones and in particular trialkoxyoctylsilanes or simethicones. In sunscreens, so-called micro- or nanopigments are preferably used. Preferably, micronized zinc oxide is used.

In addition to the two groups of primary light stabilizers mentioned above, it is also possible to use secondary light stabilizers of the antioxidant type which interrupt the photochemical reaction chain which is triggered when UN radiation penetrates into the skin. Typical examples thereof are amino acids (eg glycine, histidine, tyrosine, tryptophan) and their derivatives, imidazoles (eg urocaninic acid) and their derivatives, peptides such as D, L-canyosine, D-camosine, L-carnosine and their derivatives (eg Anserine), carotenoids, carotenes (eg α-carotene, β-carotene, lycopene) and their derivatives, chlorogenic acid and its derivatives, lipoic acid and derivatives thereof (eg dihydrolipoic acid), aurothioglucose, propylthiouracil and other thiols (eg thioredoxin, Glutathione, cysteine, cystine, cystamine and their glycosyl, Ν-acetyl, methyl, ethyl, propyl, amyl, butyl and lauryl, palmitoyl, oleyl, γ-linoleyl, cholesteryl and glyceryl esters ) and their salts, dilauryl thiodipropionate, distearyl thiodipropionate, thiodipropionic acid and derivatives thereof (esters, ethers, peptides, lipids, nucleotides, nucleosides and salts) and sulfoximine compounds (eg buthionine sulfoximines, homocysteine sulfoximine, bithine sulfones, penta, hexa, heptathionine sulfoximine) in a lot low tolerated dosages (eg pmol to μmol / kg), furthermore (metal) chelators (eg α-hydroxy fatty acids, palmitic acid, phytic acid, lactoferrin), α-hydroxy acids (eg citric acid, lactic acid, malic acid), humic acid, bile acid, bile extracts, bilirubin , Biliverdin, EDTA, EGTA and their derivatives, unsaturated fatty acids and their derivatives (eg γ-linolenic acid, linoleic acid, oleic acid), folic acid and its derivatives, ubiquinone and ubiquinol and their derivatives, vitamin C and derivatives (eg ascorbyl palmitate, magnesium). Ascorbyl phosphate, ascorbyl acetate), tocopherols and derivatives (eg vitamin E acetate), vitamin A and derivatives (vitamin A palmitate), as well as benzoic acid coniferyl benzoate, rutinic acid and its derivatives, glycosyl rutin, ferulic acid, furfurylidene glucitol, carnosine, butyl hydroxy - toluene, butylhydroxyanisole, nordihydroguaiacetic acid, nordihydroguaiaretic acid, trihydroxybutyrophenone, uric acid and derivatives thereof, mannose and derivatives thereof, superoxide dismutase, zinc and its derivatives (eg ZnO, ZnSO ₄ ) selenium and its derivatives (eg selenium methionine), Stilbenes and their derivatives (eg stilbene oxide, trans-stilbene oxide) and the inventively suitable derivatives (salts, esters, ethers, sugars, nucleotides, nucleosides, peptides and lipids) of these drugs.

Examples

Example 1: Plectin Expression

reagents:

Monoclonal Antibodies anti-plectin and secondary antibodies to fluorescein isothiocyanate FITC-conjugated anti-mouse antibodies IgG were obtained from Tebu and Clinics. PBS (phosphate buffered saline pH 7.2) and Evans blue were purchased from BioMerieux and TGF is from Sigma.

EKLN reconstructed human skin:

Reconstructed human skin of Episkin (Ekin kit) consists of a dermal support shaped from collagen. The keratinocytes were removed from this support. After removal of the keratinocytes and differentiation in an air-exposed culture, one equivalent to human skin could be obtained.

Test substances

TGF beta was used as a positive control.

The test substance used was a preparation consisting of: propylene glycol, Ruscus aculeatus root extract, Centella asiatica extract, panthenol, water, Calendula officinalis flower extract, hydrolyzed yeast proteins, Aesculus Hippocastanum extract extract, and ammonium glycyrrhizate in varying proportions by weight.

The reconstituted human skin was topically treated daily for three days with 0.01% of this preparation or with 10 ng ml TGF beta. This corresponds to a total amount of preparation of 100 microliters per reconstructed skin. After this treatment, biopsies were taken immediately and stored in liquid nitrogen until evaluated.

immunohistochemistry

Ten milimeters of cryostat section (liquid nitrogen stored biopsy) were fixed on glass slides and stored in cold acetone for ten minutes. Subsequently, the section was washed in PBS and dried in air. ^•

The section was incubated for one hour at room temperature with the monoclonal antibody antiplectin in a 1/150 solution. After washing with PBS, the section was incubated with fluorescein isothiocyanate (FITC) -conjugated anti-mouse antibody for 45 min in a 1/40 solution. Negative controls were obtained by omission of the first antibody. After extensive washing with PBS, the immuno-treated sections were incubated for 10 min. treated with Evans blue. The sections were examined with a concave laser microscope from the Finn Zeiss. quantification

The images obtained by the convocal laser microscope were converted and analyzed by mathematical morphological software (Quantimet Q500, Leica). The results were presented as percent of the reconstructed skin surface evidenced by plectin (FITC).

Results

Without treatment only a low expression of the DEJ component plectin could be detected in the reconstructed skin.

The topical treatment with the preparation of the test substances have shown a strong increase in the expression of plectin in the reconstructed skin. The positive control TGF also showed an increase in expression of plectin in the reconstructed skin.

Example 2: Perlecan expression

reagents:

Monoclonal Antibodies Anti-perlecan and secondary antibodies to fluorescein isothiocyanate FITC-conjugated anti-mouse antibody IgG were obtained from Tebu and Clinisciences. PBS (Phosphate buffered saline pH 7.2) and Evans blue were purchased from BioMereux and TGF was purchased from Sigma.

Human primary fibroblasts culture

A cell suspension of human fibroblasts was prepared by standard digestion of collagenase from adult human Dennis obtained by plastic surgery. The fibroblasts were grown on glass dishes with incubation chambers and grew in the culture medium to confluency.

Test substances

TGF beta was used as a positive control.

The test substance was a hydrolyzed Hibiscus Esculentus extract. The human fibroblasts were cultured in the presence of 0.1% of the test substance or in the presence of 10 ng / ml TGF beta in the culture medium for 6 days. Subsequently, the Perlecan expression was evaluated by iminunocytochemistry.

hmnunohistochemie

The fibroblast culture in the glass dishes was fixed in cold methanol for 10 min and washed with PBS. Subsequently, the fibroblast cultures were incubated for one hour at 37 ° C with the monoclonal antibody anti-plectin in a 1/150 solution. After washing with PBS, the section was incubated with fluorescein isothiocyanate (FITC) -conjugated anti-mouse antibody for 45 min in a 1/40 solution. Negative controls were obtained by omission of the first antibody. After extensive washing with PBS, the immuno-treated sections were incubated for 10 min. treated with Evans blue. The sections were examined with a convocal laser microscope from Zeiss.

quantification

The images obtained by the convocal laser microscope were converted and analyzed by mathematical morphological software (Quantimet Q500, Leica). The results were presented as percent of the area of fibroblast culture evidenced by perlecan (FITC).

Results

Without treatment, only a small expression of the DEJ component perlecan could be detected in the fibroblasts culture.

Treatment with the test substance has shown an increase in the expression of pearlcanin in the fibroblast culture. The positive control TGF also showed an increase in expression of periancan in the fibroblast culture.

The results from Example 1 and Example 2 show that the test substances (products of Laboratoires serobiologiques) can increase the expression of plectin and pearlcan.

Claims

claims

1. A process for the cosmetic treatment for the improvement and / or protection of dermal-epidermal junctions of the skin, scalp and mucous membrane, which comprises preparing a preparation containing at least one substance which has a modulation of plectin / HDI and / or entactin / Nidogen and / or Perlecan effected, topically applied.

2. Use of a substance which effects a modulation of plectin / HDI and / or entactin / nidogen and / or pearlcan, for the production of cosmetic agents for improving and / or protecting the dermal-epidermal junctions of the skin, scalp and mucous membrane.

3. Use of a substance which effects a modulation of plectin / HDI and / or entactin / nidogen and / or pearlcane, for the production of cosmetic agents for the protection of the skin against aging.

4. Use of a substance which effects a modulation of plectin / HDI and / or entactin / nidogen and / or perlecan, for the production of cosmetic agents for protecting the skin, scalp and mucous membrane against toxic environmental influences.

5. Use of a substance which causes a modulation of plectin / HDI and / or entactin / nidogen and / or perlecan, for the preparation of cosmetic agents zxxm protection of the skin, scalp and mucous membrane against UV light.

6. Use of a substance which effects a modulation of plectin / HDI and / or entactin / nidogen and / or perlecan, for the production of cosmetic preparations against oxidative stress.

7. Use of a substance which effects a modulation of plectin / HDI and / or entactin / nidogen and / or perlecan, for the preparation of cosmetic products against hair loss and for improving the hair properties.

8. Use according to any one of claims 2 to 7, characterized in that the substance is a plant extract, which causes a modulation of Plectin / HDI and / or Entactin / Nidogen and / or Perlecan.

9. Use according to claim 8, characterized in that the extract from the plant Pisum sativum, Ruscus Aculeatus, Centella asiatica, Calendula officinalis, Aesculus Hippocastanum, and / or Hibiscus esculentus is obtained as.

10. Use according to any one of claims 2 to 7, characterized in that the substance is selected from the group formed by mannitol, Cyclodextrin, yeast extract, panthenol, propylene glycol, ammonium glycinehizate and disodium succinate, low molecular weight peptides.

11. Cosmetic preparations containing at least one substance which effects a modulation of plectin / HDI and / or entactin / nidogen and / or perlecan, and UV sun protection factors and / or antioxidants.